title: Chitinase activity on amorphous chitin thin films: a quartz crystal microbalance with dissipation monitoring and atomic force microscopy study.
abstract: Chitinases are widely distributed in nature and have wide-ranging pharmaceutical and biotechnological applications. This work highlights a real-time and label-free method to assay Chitinase activity via a quartz crystal microbalance with dissipation monitoring (QCM-D) and atomic force microscopy (AFM). The chitin substrate was prepared by spincoating a trimethylsilyl chitin solution onto a silica substrate, followed by regeneration to amorphous chitin (RChi). The QCM-D and AFM results clearly showed that the hydrolysis rate of RChi films increased as Chitinase (from Streptomyces griseus) concentrations increased, and the optimal temperature and pH for Chitinase activity were around 37 °C and 6-8, respectively. The Chitinase showed greater activity on chitin substrates, having a high degree of acetylation, than on chitosan substrates, having a low degree of acetylation.
authors.affiliation: Department of Chemistry, Virginia Tech, Blacksburg, 24061, USA.
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Kittle
authors.forename: Chen
authors.initials: C
authors.lastname: Qian
authors.forename: Maren
authors.initials: M
authors.lastname: Roman
authors.forename: Alan R
authors.initials: AR
authors.lastname: Esker
mesh_terms: D000107:Acetylation; D001426:Bacterial Proteins / Q000737:chemistry*; D002686:Chitin / Q000737:chemistry*; D002688:Chitinases / Q000737:chemistry*; D057075:Enzyme Assays; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D018625:Microscopy, Atomic Force; D011499:Protein Processing, Post-Translational; D058525:Quartz Crystal Microbalance Techniques; D013305:Streptomyces griseus / Q000201:enzymology; D013499:Surface Properties; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D002686:Chitin; D002688:Chitinases